Polymer nanoparticle as a delivery system for ribavirin: Do nanoparticle avoid uptake by Red Blood Cells?

An investigation of rhinovirus infection on cellular uptake of poly (glycerol-adipate) nanoparticles

“…RBITC PGA NPs were prepared by interfacial deposition and fluorescent dye loading was achieved as previously reported ( Abo-zeid et al, 2018b , Abo-zeid and Garnett, 2020 ). The particle size was 110 ± 30 nm (diameter mean ± SD) and the polydispersity index 0.01, indicating a monodisperse sample.…”

“…The encapsulation efficiency and dye loading were 54 ± 13% and 0.54 ± 0.13%, respectively. This dye loading is sufficient enough to track NPs uptake by cells using flow cytometry ( Abo-zeid et al, 2018b , Abo-zeid and Garnett, 2020 ).…”

“…This study explored four different models of virally infected cells to broaden our understanding into the effect of infection on NPs uptake. We prepared fluorescently labelled RBITC PGA NPs with a dye loading that is sufficient to track the uptake of NPs by flow cytometry ( Abo-zeid et al, 2018b , Abo-zeid and Garnett, 2020 ). These NPs were used for several reasons: they are formulated from PGA, a biodegradable and biocompatible polymer with very low cytotoxic properties ( Abo-zeid and Garnett, 2020 , Zhang et al, 2014 , Kallinteri et al, 2005 ;), and RBITC is retained in the NPs for a prolonged period of time ( Meng et al, 2006 ), allowing us to track the uptake of NPs rather than free dye ( Abo-zeid et al, 2018b ).…”

“…We prepared fluorescently labelled RBITC PGA NPs with a dye loading that is sufficient to track the uptake of NPs by flow cytometry ( Abo-zeid et al, 2018b , Abo-zeid and Garnett, 2020 ). These NPs were used for several reasons: they are formulated from PGA, a biodegradable and biocompatible polymer with very low cytotoxic properties ( Abo-zeid and Garnett, 2020 , Zhang et al, 2014 , Kallinteri et al, 2005 ;), and RBITC is retained in the NPs for a prolonged period of time ( Meng et al, 2006 ), allowing us to track the uptake of NPs rather than free dye ( Abo-zeid et al, 2018b ). RBITC dye gives a good fluorescence in the acidic pH of lysosomal compartment ( Garnett and Baldwin, 1986 ) leading to effective detection of NPs taken up by cells using flow cytometry ( Abo-zeid and Garnett, 2020 , Abo-zeid et al, 2018b ).…”

“…RBITC PGA NPs were prepared and fluorescent dye loading was optimized as previously reported ( Abo-zeid et al, 2018b , Abo-zeid and Garnett, 2020 ). Briefly, RBITC (200 μl, 1 mg/ml, in methanol) was added into an aqueous phase (HEPES buffer, 10 mM, pH 7.4, 7 ml).…”